scholarly journals Molecular Characterisation of Soil-Dwelling Bacillus thuringiensis using Transcriptional Regulator, XRE Gene and the Crystal Protein, cry2 gene

2021 ◽  
Vol 6 (1) ◽  
pp. 153-159
Author(s):  
E.O Akinyelure ◽  
◽  
D.A. Machido ◽  
H. I. Atta
Keyword(s):  
Bacillus Thuringiensis ◽  
Environmental Samples ◽  
Plant Pathogens ◽  
Transcriptional Regulator ◽  
Soil Samples ◽  
Molecular Techniques ◽  
Crystal Protein ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Sporulation Phase

Bacillus thuringiensis (Bt) is the organism that is used most frequently in biological pest management, which is distinguished by the capacity to possess crystalline inclusions throughout the sporulation phase. There is an increasing need to use biological control in controlling plant pathogens due to the inherent advantages. However, the detection of Bt has become more time consuming and cumbersome due to the numerous available crystal genes. The goal of the study was to isolate strains of Bacillus thuringiensis from the soil, characterise the isolates using the transcriptional regulator, XRE gene and the crystal proteins cry2gene and compare the efficiency of these two biomarkers in identifying Bt species. Five different Bacillus thuringiensis strains were isolated from soil samples in Zaria, Nigeria. Polymerase chain reaction was used to detect the existence of the cry2 and XRE genes. Four (80%) of the five isolates harboured the XRE genes, while none (0%) harboured the cry2 genes. This observation is a likely indication that the XRE gene is a reliable biomarker in the identification of Bt isolates from environmental samples. In order to ensure speed and reproducibility in the detection of Bt from environmental samples, molecular techniques targeting the XREgene are recommended. Keywords: Bacillus thuringiensis; transcriptional regulator, XRE; crystal protein, cry2

scholarly journals Diversity ofLeptospiraspp. in Rats and Environment from Urban Areas of Sarawak, Malaysia

2017 ◽  
Vol 2017 ◽  
pp. 1-8 ◽  
Author(s):  
Chai Fung Pui ◽  
Lesley Maurice Bilung ◽  
Kasing Apun ◽  
Lela Su’ut
Keyword(s):  
Polymerase Chain Reaction ◽  
Water Samples ◽  
Urban Areas ◽  
Environmental Samples ◽  
Soil Samples ◽  
Chain Reaction ◽  
Prevalence Studies ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Soil And Water

Various prevalence studies onLeptospirain animals and humans, as well as environmental samples, had been conducted worldwide, including Malaysia. However, limited studies have been documented on the presence of pathogenic, intermediate, and saprophyticLeptospirain selected animals and environments. This study was therefore conducted to detectLeptospiraspp. in rats, soil, and water from urban areas of Sarawak using the polymerase chain reaction (PCR) method. A total of 107 rats, 292 soil samples, and 324 water samples were collected from April 2014 to February 2015. PathogenicLeptospirawas present in 5.6% (6/107) of rats, 11.6% (34/292) of soil samples, and 1.9% (6/324) of water samples. IntermediateLeptospirawas present in 2.7% (8/292) of soil samples and 1.9% (6/324) of water samples. SaprophyticLeptospirawas present in 10.3% (11/107) of rats, 1.4% (4/292) of soil samples, and 0.3% (1/324) of water samples. From this study, 76Leptospiraspp. were isolated. Based on DNA sequencing, the dominantLeptospiraspp. circulating in urban areas of Sarawak are pathogenicLeptospira noguchii, intermediateLeptospira wolffiiserovar Khorat, and saprophyticLeptospira meyeri, respectively. Overall, this study provided important surveillance data on the prevalence ofLeptospiraspp. from rats and the environment, with dominant local serovars in urban areas of Sarawak.

Extraction of microsporidial DNA from modified trichrome-stained clinical slides and subsequent species identification using PCR sequencing

Parasitology ◽  
2008 ◽  
Vol 135 (6) ◽  
pp. 701-703 ◽  
Author(s):  
K. S. CHAN ◽  
T. H. KOH
Keyword(s):  
Polymerase Chain Reaction ◽  
Species Identification ◽  
Environmental Samples ◽  
Species Level ◽  
Molecular Techniques ◽  
Chain Reaction ◽  
Clinical Specimens ◽  
Polymerase Chain ◽  
Pcr Conditions ◽  
Hiv Negative

SUMMARYMolecular techniques involving polymerase chain reaction (PCR) and sequencing provide a relatively simple and objective means of identifying microsporidia to species level. We modified previously described methods of DNA extraction and PCR conditions for identification of microsporidia from museum slides, clinical specimens and environmental samples and successfully identifiedVittaforma corneaein 11 out of 13 cases of microsporidial infection from used trichrome-stained slides of corneal scrapings from HIV-negative patients with keratoconjunctivitis.

Limitations of Molecular Biological Techniques for Assessing the Virological Safety of Foods†

1999 ◽  
Vol 62 (6) ◽  
pp. 691-697 ◽  
Author(s):  
GARY P. RICHARDS
Keyword(s):  
Polymerase Chain Reaction ◽  
Environmental Samples ◽  
Hepatitis A ◽  
Enteric Viruses ◽  
Molecular Techniques ◽  
Technical Expertise ◽  
Chain Reaction ◽  
Assay Sensitivity ◽  
Testing Laboratories ◽  
Polymerase Chain

Enteric viruses, including hepatitis A, Norwalk, and Snow Mountain viruses, Hawaii agent, and rotaviruses have been associated with outbreaks of foodborne illness. Classical culturing procedures are available for poliovirus; however, hepatitis A, Norwalk, and many of the other viruses and agents cannot be propagated in cell culture, therefore, molecular biological tools have emerged as a possible means to detect enteric viruses in foods and environmental samples. There are limitations however in the application of polymerase chain reaction and reverse transcription polymerase chain reaction that restrict their usefulness for measuring the virological safety of foods. The most serious limitation is that molecular techniques fail to discriminate between viable and inactivated viruses even though inactivated viruses pose no threat to the consumer and may be present at levels substantially higher than the virulent forms. Other disadvantages include a lack of assay sensitivity and specificity, high assay costs, and a level of technical expertise not available in most food-testing laboratories. Overall, scientific advances in the development of molecular biological tools have outpaced the demonstration of their validity in assessing the virological safety of foods.

Polymerase chain reaction amplification and typing of rotavirus in environmental samples

1995 ◽  
Vol 31 (5-6) ◽  
pp. 371-374 ◽  
Author(s):  
R. Gajardo ◽  
R. M. Pintó ◽  
A. Bosch
Keyword(s):  
Polymerase Chain Reaction ◽  
Environmental Samples ◽  
Polymerase Chain Reaction Amplification ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Group A ◽  
Reaction Amplification ◽  
Stool Specimens ◽  
Polymerase Chain

A reverse transcription polymerase chain reaction (RT-PCR) assay is described that has been developed for the detection and serotyping of group A rotavirus in stool specimens and concentrated and non-concentrated sewage specimens.

scholarly journals Assessment of the Nutraceutical Effects of Oleuropein and the Cytotoxic Effects of Adriamycin, When Administered Alone and in Combination, in MG-63 Human Osteosarcoma Cells

Nutrients ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 354
Author(s):  
Katerina Gioti ◽  
Anastasia Papachristodoulou ◽  
Dimitra Benaki ◽  
Nektarios Aligiannis ◽  
Alexios-Leandros Skaltsounis ◽  
...  
Keyword(s):  
Chemotherapeutic Agent ◽  
Molecular Techniques ◽  
Osteosarcoma Cells ◽  
Elisa Method ◽  
Chain Reaction ◽  
Cytotoxic Effects ◽  
Human Osteosarcoma ◽  
Human Osteosarcoma Cells ◽  
Anti Cancer ◽  
Polymerase Chain

Oleuropein (OLEU) is the most distinguished phenolic compound found in olive fruit and the leaves of Olea europaea L., with several pharmacological properties, including anti-cancer actions. Adriamycin (ADR) is an anthracycline widely used as a chemotherapeutic agent, although it presents significant side effects. The aim of the present study was to investigate the effect of oleuropein alone (20 μg/mL) and in co-treatment with ADR (50 nM), in MG-63 human osteosarcoma cells. Therefore, cellular and molecular techniques, such as MTT assay, flow cytometry, real-time Polymerase Chain Reaction (PCR), western blot and Elisa method, as well as Nuclear Magnetic Resonance (NMR) spectroscopy, were applied to unveil changes in the signal transduction pathways involved in osteosarcoma cells survival. The observed alterations in gene, protein and metabolite levels denote that OLEU not only inhibits MG-63 cells proliferation and potentiates ADR’s cytotoxicity, but also exerts its action, at least in part, through the induction of autophagy.

Diagnostic Accuracy of a Direct Panfungal Polymerase Chain Reaction Assay Performed on Stained Cytology Slides

2021 ◽  
pp. 030098582199156
Author(s):  
Alexandra N. Myers ◽  
Unity Jeffery ◽  
Zachary G. Seyler ◽  
Sara D. Lawhon ◽  
Aline Rodrigues Hoffmann
Keyword(s):  
Polymerase Chain Reaction ◽  
Diagnostic Accuracy ◽  
Molecular Techniques ◽  
Fungal Culture ◽  
Chain Reaction ◽  
Identification Of Fungi ◽  
Fungal Dna ◽  
Polymerase Chain ◽  
Panfungal Pcr ◽  
Pcr Targeting

Molecular techniques are increasingly being applied to stained cytology slides for the diagnosis of neoplastic and infectious diseases. Such techniques for the identification of fungi from stained cytology slides have not yet been evaluated. This study aimed to assess the diagnostic accuracy of direct (without nucleic acid isolation) panfungal polymerase chain reaction (PCR) followed by sequencing for identification of fungi and oomycetes on stained cytology slides from dogs, cats, horses, and other species. Thirty-six cases were identified with cytologically identifiable fungi/oomycetes and concurrent identification via fungal culture or immunoassay. Twenty-nine controls were identified with no cytologically or histologically visible organisms and a concurrent negative fungal culture. Direct PCR targeting the internal transcribed spacer region followed by sequencing was performed on one cytology slide from each case and control, and the sensitivity and specificity of the assay were calculated. The sensitivity of the panfungal PCR assay performed on stained cytology slides was 67% overall, 73% excluding cases with oomycetes, and 86% when considering only slides with abundant fungi. The specificity was 62%, which was attributed to amplification of fungal DNA from control slides with no visible fungus and negative culture results. Direct panfungal PCR is capable of providing genus- or species-level identification of fungi from stained cytology slides. Given the potential of panfungal PCR to amplify contaminant fungal DNA, this assay should be performed on slides with visible fungi and interpreted in conjunction with morphologic assessment by a clinical pathologist.

scholarly journals Myostatin Gene Expression and Its Application on Goat Breeding Programme

2018 ◽  
Vol 27 (2) ◽  
pp. 89 ◽  
Author(s):  
Aron Batubara
Keyword(s):  
Muscle Hypertrophy ◽  
Body Weight Gain ◽  
Muscle Growth ◽  
Single Strand Conformation Polymorphism ◽  
Molecular Techniques ◽  
Chain Reaction ◽  
Myostatin Gene ◽  
Production Parameters ◽  
Polymerase Chain ◽  
Conformation Polymorphism

Characteristics of double muscled growth in animals are influenced by myostatin gene (MSTN). Myostatin gene is known as a member of the growth gene's superfamily (TGF-β) which works to suppress the muscle growth. However, the presence of six mutations on MSTN cause the gene inactive, and trigger the occurrence of muscle hypertrophy. Identification of myostatin gene was conducted by molecular techniques, and the most common method is polymerase chain reaction followed by single-strand conformation polymorphism (PCR-SSCP). Research on sheep and goat in several countries showed that there had been several variations occurred in myostatin gene but further studies are required to correlate these variations to body weight gain and other important production parameters. For goat production in Indonesia, myostatin mutations cause double muscling that can be utilised for genetic improvement in goat breeding plan to produce a new breed with high quality meat.

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